Figure 9.
β-catenin targets vinculin to AJs and centrioles. (A–C) “En face” pictures of chicken NTs transfected at stage HH12 for 24 h with Cep152-GFP (yellow) and VCL·mCherry (A) or mCherry (B) or VCLA50I·mCherry (magenta) and stained with anti N-cadherin antibody (green). The magenta channel is shown alone in right-handed panels for clarity. The position of representative AJs and centrosomes are indicated by arrowheads and arrows, respectively. The images are Z projections of the confocal planes containing the AJs and the centrioles (7 to 10 planes spaced 0.15 µm). Scale bar = 2 μm. (D) Scheme of the DNA vector used in this figure. (E–J) Images of transverse sections of chicken NTs transfected at stage HH12 for 24 h with shControl or shVCL plus empty vector (pCIG), VCL-WT, or VCL-A50I, stained with phalloidin (actin, gray scale), the GFP expressed in the transfected halves of the NTs is shown at the right hand of each image to show transfection level. Scale bar = 50 μm. (K) Dot plot showing the apicobasal position of the nuclei from the NTs shown in Fig. 9 E. The apical–basal position of 500 nuclei was measured for each condition. The black bars indicate the median. One-way ANOVA plus Dunn’s multiple comparisons test. n = 10 sections per condition from three experiments using at least three embryos per condition. (L) Bar graph showing the percentage of mitotic (PH3+) cells relative to transfected cells (GFP+) in chicken NTs electroporated at stage HH12 for 24 h with shControl or shVCL plus empty vector (pCIG), VCL-WT, or VCL-A50I. Mean ± SEM. One-way ANOVA plus Tukey’s multicomparison test, n = 11–16 sections from two experiments using at least three embryos per condition. (M–R) Flow cytometry analysis profiles showing DNA content (Hoechst fluorescence intensity) and cell number of GFP+ population from chicken NTs transfected at stage HH12 for 24 h with shControl or shVCL plus empty vector (pCIG), VCL-WT, or VCL-A50I. (S) Bar graph showing the percentage of cells in each cell cycle phase from the profiles shown in Fig. 9, M–R. Mean ± SEM. Two-way ANOVA plus Tukey’s multicomparison test. Two experiments using at least three embryos per condition. A minimum of 2,000 cells per condition were analyzed in each experiment. ns = non-significant, * = P < 0.05, ** = P < 0.01, *** = P < 0.001, **** = P < 0.0001.

β-catenin targets vinculin to AJs and centrioles. (A–C) “En face” pictures of chicken NTs transfected at stage HH12 for 24 h with Cep152-GFP (yellow) and VCL·mCherry (A) or mCherry (B) or VCLA50I·mCherry (magenta) and stained with anti N-cadherin antibody (green). The magenta channel is shown alone in right-handed panels for clarity. The position of representative AJs and centrosomes are indicated by arrowheads and arrows, respectively. The images are Z projections of the confocal planes containing the AJs and the centrioles (7 to 10 planes spaced 0.15 µm). Scale bar = 2 μm. (D) Scheme of the DNA vector used in this figure. (E–J) Images of transverse sections of chicken NTs transfected at stage HH12 for 24 h with shControl or shVCL plus empty vector (pCIG), VCL-WT, or VCL-A50I, stained with phalloidin (actin, gray scale), the GFP expressed in the transfected halves of the NTs is shown at the right hand of each image to show transfection level. Scale bar = 50 μm. (K) Dot plot showing the apicobasal position of the nuclei from the NTs shown in Fig. 9 E. The apical–basal position of 500 nuclei was measured for each condition. The black bars indicate the median. One-way ANOVA plus Dunn’s multiple comparisons test. n = 10 sections per condition from three experiments using at least three embryos per condition. (L) Bar graph showing the percentage of mitotic (PH3+) cells relative to transfected cells (GFP+) in chicken NTs electroporated at stage HH12 for 24 h with shControl or shVCL plus empty vector (pCIG), VCL-WT, or VCL-A50I. Mean ± SEM. One-way ANOVA plus Tukey’s multicomparison test, n = 11–16 sections from two experiments using at least three embryos per condition. (M–R) Flow cytometry analysis profiles showing DNA content (Hoechst fluorescence intensity) and cell number of GFP+ population from chicken NTs transfected at stage HH12 for 24 h with shControl or shVCL plus empty vector (pCIG), VCL-WT, or VCL-A50I. (S) Bar graph showing the percentage of cells in each cell cycle phase from the profiles shown in Fig. 9, M–R. Mean ± SEM. Two-way ANOVA plus Tukey’s multicomparison test. Two experiments using at least three embryos per condition. A minimum of 2,000 cells per condition were analyzed in each experiment. ns = non-significant, * = P < 0.05, ** = P < 0.01, *** = P < 0.001, **** = P < 0.0001.

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